体内胚和体细胞克隆胚受体牛妊娠不同阶段血液中主要生殖激素含量变化的研究

发布时间：2018-03-03 08:01

本文选题：体内胚　切入点：体细胞克隆胚　出处：《内蒙古农业大学》2015年硕士论文　论文类型：学位论文

【摘要】：本实验通过对体细胞克隆胚受体牛和体内胚受体牛,在其妊娠不同阶段对血液中主要生殖激素含量变化的研究,试图从受体母牛生殖激素水平方而解释体细胞克隆牛异常妊娠的原因,预测牛胎儿流产,从而提高体细胞克隆牛生产效率。构建一个尽可能完整的常规受孕母牛在妊娠不同阶段的主要生殖激素水平参考指标,为体细胞克隆牛生产提供可靠的参考指标,在已有的基础上优化一个更好的体细胞克隆牛生产管理体系。使用加拿大进口FSH,采用CIDR+FSH+PGF2a法对供体和牛进行超排,从而获得体内胚。采用组织块贴壁法进行和牛胎儿成纤维细胞的原代培养,选择P5代细胞作为核移植供体细胞,通过电融合法获得体细胞克隆重构胚选择合适受体牛进行体内胚和体细胞克隆胚的移植,在体内胚和体细胞克隆胚受体牛妊娠不同阶段,通过化学发光免疫分析法(CLIA)对其血液中主要生殖激素(FSH、LH、和P)的含量进行测定。使用加拿大进口FSH的总剂量为380-420IU,成功获得58枚可用和牛体内胚。成功建立了和牛胎儿成纤维细胞系,构建了和牛体细胞克隆胚,总共38枚。建立了体内胚受体牛在妊娠不同阶段血液中生殖激素(FSH、LH、E2、P)含量变化的参考指标,体细胞克隆胚受体牛与该指标做了比较,在生殖激素水平分析了体细胞克隆胚受体牛流产的原因。
[Abstract]:In this study, we studied the changes of the main reproductive hormones in the blood of somatic cloned embryo receptor cattle and in vivo embryo receptor cattle at different stages of pregnancy. This paper attempts to explain the causes of abnormal pregnancy of somatic cloned cattle from the reproductive hormone level of recipient cows, and to predict the abortion of bovine fetuses. In order to improve the productivity of somatic cloned cattle, to construct a complete reference index of reproductive hormone level of conventional pregnant cows in different stages of pregnancy, and to provide reliable reference index for the production of somatic cloned cattle. A better system of production management of somatic cloned cattle was optimized on the basis of existing methods. The superovulation of donor and cattle was carried out by CIDR FSH PGF2a with imported FSHs from Canada. In vivo embryos were obtained. Primary culture of bovine fetal fibroblasts was carried out by tissue mass adherence method. P5 passage cells were selected as nuclear transfer donor cells. Somatic clone reconstructed embryos were obtained by electrofusion to select suitable recipient bovine embryos for in vivo and somatic cell clone embryos transplantation. In vivo embryos and somatic cell cloned embryos receptor cattle pregnancy stage. Chemiluminescence immunoassay (CLIA) was used to determine the contents of the main reproductive hormones FSHLH, and Pin in the blood. The total dose of imported FSH from Canada was 380-420 IUUs, 58 usable and in vivo embryos were successfully obtained. Infantile fibroblast cell line, A total of 38 somatic cloned embryos were constructed. A reference index was established for the change of the content of reproductive hormone (FSHLHN E2P) in the blood of somatic receptor cattle in different stages of pregnancy. The somatic cloned embryo receptor cattle were compared with this index. The causes of bovine abortion of somatic cloned embryo receptor were analyzed at the reproductive hormone level.
【学位授予单位】：内蒙古农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S823;S857.2

【参考文献】